Functional characterization of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D)

During the last few decades, starch-degrading enzymes have emerged as promising tools for transforming biomass. Nevertheless, their functional characterization, based essentially on colorimetric tests, is still fastidious and time-consuming. In order to overcome these hurdles, we have developed a te...

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Bibliographic Details
Published in:Sensors and actuators. B, Chemical Chemical, 2013-01, Vol.176, p.1038-1043
Main Authors: Bouchet-Spinelli, Aurélie, Coche-Guérente, Liliane, Armand, Sylvie, Lenouvel, François, Labbé, Pierre, Fort, Sébastien
Format: Article
Language:English
Subjects:
biomass
colorimetry
Glycoside hydrolases
hydrolysis
monitoring
mutants
quartz
Quartz crystal microbalance with dissipation monitoring
Real-time label-free detection
Starch degradation
transferases
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Summary:During the last few decades, starch-degrading enzymes have emerged as promising tools for transforming biomass. Nevertheless, their functional characterization, based essentially on colorimetric tests, is still fastidious and time-consuming. In order to overcome these hurdles, we have developed a test for real-time and label-free detection of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D). By starch-functionalization of surfaces, we were able to discriminate clearly enzymes with different hydrolysis patterns (endo versus exo). The variation of the hydrolytic activity between the wild-type and a mutant of a cyclodextrin glucosyl transferases could also be monitored acoustically. The high sensitivity reached (down to 6.5nkat/L) opens the prospect of using fewer enzyme quantities for functional tests and is a further advantage for this easily used and efficient tool for enzyme characterization.
ISSN:0925-4005
1873-3077
DOI:10.1016/j.snb.2012.10.013